Preparation method of mellea armillaria sporophore antioxidant peptide
A technology of antioxidant peptides and hazel mushrooms, which is applied to the preparation method of peptides, chemical instruments and methods, peptides, etc., can solve the problem that the antioxidant activity of proteins in hazel mushrooms cannot be known, and achieve the effect of improving the extraction rate
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Embodiment 1
[0037] A preparation method of hazelnut mushroom antioxidant peptide, extracting protein according to the following steps:
[0038] (1) Wash the hazel mushrooms, dry them, pulverize them and pass through a 60 mesh sieve, add them into distilled water according to the material-to-liquid ratio of 1:45, after mixing evenly, adjust the pH to 10 with 0.1 mol / L NaOH, and at 80°C Water bath for 1.5h;
[0039] (2) after the alkali extraction is completed, filter out the residue, add distilled water according to the ratio of material to liquid of 1:45, after mixing evenly, add 0.1mol / L NaOH to adjust pH to 9, and ultrasonically treat with 200W power at room temperature for 14-16min;
[0040] (3) Adjust the pH of the supernatant after extraction to 3.7 with 0.1 mol / L hydrochloric acid, then place it at 4°C for 12 hours, and then centrifuge at 10,000 rpm for 20 minutes at this temperature to obtain protein 1, and take the isoelectric point The supernatant after precipitation was added w...
Embodiment 2
[0060] A preparation method of hazelnut mushroom antioxidant peptide is carried out according to the following steps:
[0061] (1) Extraction of protein
[0062] (1) Wash the hazel mushrooms, dry them, pulverize them and pass through a 60 mesh sieve, add them into distilled water according to the material-to-liquid ratio of 1:45, after mixing evenly, adjust the pH to 10 with 0.1 mol / L NaOH, and at 80°C Water bath for 1.5h;
[0063] (2) after the alkali extraction is completed, filter out the residue, add distilled water according to the ratio of material to liquid of 1:45, after mixing evenly, add 0.1mol / L NaOH to adjust pH to 9, and ultrasonically treat with 200W power at room temperature for 14-16min;
[0064] (3) Adjust the pH of the supernatant after the extraction to 3.6 with 0.1 mol / L hydrochloric acid, then place it at 4°C for 12 hours, and then centrifuge at 10,000 rpm for 20 minutes at this temperature to obtain protein 1, and take the isoelectric point The supernat...
Embodiment 3
[0079] Gel Filtration Chromatography:
[0080] Take an appropriate amount of gel Sephadex G-25, add excess distilled water to soak for 24h at room temperature, stir continuously until fully swollen, and pour off the upper floating material. The swollen gel was poured into the column along the inner wall of the column at one time, without bubbles and layers. Equilibrate the column bed with pure water until the baseline is stable. Antioxidant peptides <3kD were prepared into a solution with a concentration of 30mg / mL, sterilized by filtration through a sterile filter, and loaded with 2mL. Elute with pure water, control the flow rate at 2-3mL / 5min, measure the OD254nm value, and collect the samples at the elution peak to obtain component A1 and component A2, which are concentrated and lyophilized.
[0081] Its antioxidant activity was determined. The result is as Image 6 Shown: Both component A1 and component A2 have antioxidant activity in a dose-dependent manner. When the...
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